2011 Fiscal Year Final Research Report
Fluorescence-signaling and imaging of oxidative stress on the basis of diverse fluorescent proteins originating from luminous bacteria
| Project/Area Number |
21370071
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| Research Category |
Grant-in-Aid for Scientific Research (B)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
Biophysics
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| Research Institution | Kyoto Institute of Technology |
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Principal Investigator |
KARATANI Hajime 京都工芸繊維大学, 工芸科学研究科, 教授 (10169659)
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| Co-Investigator(Kenkyū-buntansha) |
KITADOKORO Kengo 京都工芸繊維大学, 工芸科学研究科, 准教授 (60283587)
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| Co-Investigator(Renkei-kenkyūsha) |
TAJIMA Kunihiko 京都工芸繊維大学, 工芸科学研究科, 教授 (50163457)
KANAORI Kenji 京都工芸繊維大学, 工芸科学研究科, 准教授 (30273543)
YAMADA Etsu 京都工芸繊維大学, 環境科学センター, 教授 (30159214)
WADA Minoru 長崎大学, 大学院・水産・環境科学総合研究科, 准教授 (70292860)
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| Project Period (FY) |
2009 – 2011
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| Keywords | 生体生命情報学 / 生物物理 / 酸化ストレス / 蛍光タンパク質 / 発光バクテリア |
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| Research Abstract |
Live-cell fluorescence imaging method of mitochondrion in a budding yeast(Saccharomyces cerevisiae(Sc)) as a model of eukaryotic cell has been established by use of recombinant redox yellow fluorescent proteins(Y1-yellow) and blue fluorescent protein(Y1-blue), originating from luminous bacterium Aliivibrio sifiae Y1, both of which exhibits the strong fluorescence only in their oxidative states. To endow such unique fluorescence properies to Sc, the genes encoding Y1-yellow, fused with a sequence targetting at mitochondrion, and Y1-blue were newly constructed. From a series of fluorescence imaging of Sc transformed with Y1-yellow gene, of which expression was designed to be localized at mitochondrial sites, it was demonstrated that Y1-yellow is profitable to visualize the mitochondrial dynamics as well as the reactive oxygen species(ROS) in living cells. Fluorescence imaging of Sc transformed with Y1-blue gene was also useful for the visualization of the respiratory activity as well as formation and extinction of ROS. Furthermore, it was demonstrated that Y1-blue is promising for the visualization of cytochrome c, which is a key factor in an intrinsic apoptosis cascade arising from the excess ROS formation in the respiratory electron transport.
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